Probiotics for use in relieving symptoms associated with gastronitestinal disorders

ABSTRACT

The present invention provides probiotic compositions suitable for relieving symptoms associated with gastrointestinal disorders. In particular, the present invention provides compositions and methods to relieve symptoms associated with functional bowel disorders, irritable bowel syndrome, functional diarrhea, functional bloating, and other symptoms.

RELATED APPLICATIONS

The present application claims priority to U.S. Prov. Pat. Appln. Ser.No. 60/979,187, entitled “Probiotics for Use in Relieving SymptomsAssociated with Gastrointestinal Disorders,” filed Oct. 11, 2007.

FIELD OF THE INVENTION

The present invention provides probiotic compositions suitable forrelieving symptoms associated with gastrointestinal disorders. Inparticular, the present invention provides compositions and methods torelieve symptoms associated with functional bowel disorders, irritablebowel syndrome, functional diarrhea, functional bloating, and othersymptoms.

BACKGROUND OF THE INVENTION

Per the Rome Foundation (McLean, Va.), the Rome II diagnostic criteriafor functional bowel disorders (FBD) refer to various symptoms localizedto the mid or lower gastrointestinal (GI) tract. FBD include numerousclinical subgroups, such as irritable bowel syndrome (IBS), functionaldiarrhea, functional constipation, functional abdominal bloating, etc.These disorders are characterized by a variable combination of chronicor recurrent GI symptoms that are not due to structural or biochemicalabnormalities in the affected patients. IBS is the most commonfunctional GI disorder, affecting a large number of adults worldwide. Asthe pathophysiology of IBS and other FBDs is not well understood andcurrently available drug regimens are very limited, there remains a needfor methods and compositions suitable for relieving the symptoms ofthese disorders.

SUMMARY OF THE INVENTION

The present invention provides probiotic compositions suitable forrelieving symptoms associated with gastrointestinal disorders. Inparticular, the present invention provides compositions and methods torelieve symptoms associated with functional bowel disorders, irritablebowel syndrome, functional diarrhea, functional bloating, and othersymptoms.

In some embodiments, the present invention provides a combination of L.acidophilus and B. animalis subsp. lactis Bi-07. In some preferredembodiments, the present invention provides a combination of L.acidophilus NCFM® (PTA-4797) and B. animalis subsp. lactis Bi-07(PTA-4802). In some further particularly preferred embodiments, thepresent invention provides dietary supplements that contain L.acidophilus NCFM® and Bifidobacterium lactis Bi-07. In some furtherpreferred embodiments, the dietary supplements containing L. acidophilusNCFM® and Bifidobacterium lactis Bi-07 are administered to subjects at adosage in a range from about 1×10⁹ CFU to about 2×10¹¹ CFU totalprobiotic bacteria/day. In some alternative embodiments the dosage isabout 2×10¹¹ CFU total probiotic bacteria/day. However, it is notintended that the invention be limited to a specific dosage, as it iscontemplated that different dosages will find use in different settingsand/or with different patients.

The present invention also provides methods and compositions for use inimproving the symptoms of bloating and distention in patients with FBD.In some preferred embodiments, the present invention providescompositions comprising L. acidophilus and B. animalis subsp. lactis andmethods for their use in relieving the symptoms of FBD. In someparticularly preferred embodiments, the present invention providescompositions comprising L. acidophilus NCFM® and B. animalis subsp.lactis Bi-07 and methods for their use in relieving the symptoms of FBD.

DESCRIPTION OF THE INVENTION

The present invention provides probiotic compositions suitable forrelieving symptoms associated with gastrointestinal disorders. Inparticular, the present invention provides compositions and methods torelieve symptoms associated with functional bowel disorders, irritablebowel syndrome, functional diarrhea, functional bloating, and othersymptoms.

In particular, the present invention provides probiotic cultures ofLactobacillus and Bifidobacterium suitable for use with human subjects.In some particularly preferred embodiments, L. acidophilus is providedin combination with B. animalis. In yet further embodiments, B. animalissubsp. lactis (also referred to as “B. lactis” herein) is provided incombination with L. acidophilus NCFM®. In still further particularlypreferred embodiments, combinations of these organisms are provided. Inyet additional embodiments, the B. animalis subsp. lactis (also referredto as “B. lactis” herein) is B. animalis subsp. lactis is the ATCCstrain designated as PTA-4802, and the L. acidophilus NCFM® is the ATCCstrain designated as PTA-4797.

During the development of the present invention, it was found thatadministration of these probiotics provided relief of GI symptoms. Thus,the present invention provides methods and compositions comprising acombination of these probiotic bacteria useful in relieving symptomsassociated with FBD. In some particularly preferred embodiments, thecompositions find use in relieving symptoms such as bloating anddistension. However, it is not intended that the present invention belimited to these specific symptoms.

Definitions

Unless otherwise indicated, the practice of the present inventioninvolves conventional techniques commonly used in food microbiology,nutritional supplements, gastrointestinal medicine, epidemiology,molecular biology, microbiology, protein purification, and industrialenzyme use and development, all of which are within the skill of theart. All patents, patent applications, articles and publicationsmentioned herein, both supra and infra, are hereby expresslyincorporated herein by reference.

Furthermore, the headings provided herein are not limitations of thevarious aspects or embodiments of the invention, which can be had byreference to the specification as a whole. Accordingly, the termsdefined immediately below are more fully defined by reference to thespecification as a whole. Nonetheless, in order to facilitateunderstanding of the invention, definitions for a number of terms areprovided below.

Unless defined otherwise herein, all technical and scientific terms usedherein have the same meaning as commonly understood by one of ordinaryskill in the art to which this invention pertains. Although any methodsand materials similar or equivalent to those described herein find usein the practice of the present invention, preferred methods andmaterials are described herein. Accordingly, the terms definedimmediately below are more fully described by reference to theSpecification as a whole. Also, as used herein, the singular terms “a,”“an,” and “the” include the plural reference unless the context clearlyindicates otherwise. It is to be understood that this invention is notlimited to the particular methodology, protocols, and reagentsdescribed, as these may vary, depending upon the context they are usedby those of skill in the art.

It is intended that every maximum numerical limitation given throughoutthis specification includes every lower numerical limitation, as if suchlower numerical limitations were expressly written herein. Every minimumnumerical limitation given throughout this specification will includeevery higher numerical limitation, as if such higher numericallimitations were expressly written herein. Every numerical range giventhroughout this specification will include every narrower numericalrange that falls within such broader numerical range, as if suchnarrower numerical ranges were all expressly written herein.

As used herein, the term “food” refers to any nutritional item thatprovides nourishment to a plant and/or animal. It is not intended thatthe term be limited to any particular item, as it is used in referenceto any substance taken into and assimilated by a plant or animal to keepit alive. It is also not intended that the term be limited to “solid”food, as liquid nourishment is encompassed by the definition. Indeed insome embodiments, liquid nourishment is preferred over solid food items.In some preferred embodiments, the term is specifically used inreference to food for human consumption.

As used herein, the term “feed” refers to any nutritional item thatprovides nourishment to non-human animals. It is not intended that theterm be limited to any particular item, as it is used in reference toany substance taken into and assimilated by a plant or animal to keep italive. It is also not intended that the term be limited to “solid” food,as liquid nourishment is encompassed by the definition. Indeed in someembodiments, liquid nourishment is preferred over solid food items.

As used herein, the terms “nutritional supplement” and “dietarysupplement” refer to any product that is added to the diet. In someparticularly preferred embodiments, nutritional supplements are taken bymouth and often contain one or more dietary ingredients, including butnot limited to vitamins, minerals, herbs, amino acids, enzymes, andcultures of organisms. In some particularly preferred embodiments, theprobiotics of the present invention are provided in the absence ofadditional bioactive compounds. As used here in “bioactive” refers tocompositions/compounds that are active within the body. Bioactivesinclude enzymes and other compositions that have various functions,including but not limited to increasing or decreasing absorption ofnutritional substances, modifying the composition of and metabolicby-products from the intestinal microbiota, modifying the expression ofthe intestinal immune system, etc.

As used herein, the term “neutraceutical” refers to a food/dietarysupplement that is believed and/or taken to provide health benefits.

As used herein, the term “probiotic” refers to a live microbial foodingredient that is beneficial to health.

As used herein, the term “prebiotic” refers to a non-digestible foodingredient that beneficially affects a human and/or other animal thatingests the prebiotic. In preferred embodiments, prebiotics selectivelystimulate the growth and/or activity of a limited number of bacterialtypes in the intestinal tract, such that the health of the human and/orother animal is improved.

As used herein, the term “synbiotic” refers to a mixture of prebioticsand probiotics.

As used herein, the terms “illness” and “disease” refer to any deviationfrom or interruption of the normal structure and/or function of any bodypart, organ, or system that is manifested by a characteristic set ofsymptoms and signs. The term encompasses conditions with known orunknown etiology and/or pathology.

As used herein, the term “treating” refers to the providing ofcompositions that result in the improvement, amelioration, and/orremedying of a disease, disorder, or symptom of disease or condition.

As used herein, the terms “oral administration,” and “per os” refer tothe taking of food and/or supplements by mouth.

As used herein, the terms “prevention of illness” and “prevention ofdisease” refer to measures taken to avoid the incidence ofillness/disease. In some embodiments, “prophylactic” measures are takenin order to avoid disease/illness.

As used herein, the term “symptom of disease” refers to any subjectiveof disease and/or a patient's condition. It is used in reference to anysuch evidence as perceived by the patient.

As used herein, the term “sign of disease” refers to an indication ofthe existence of disease/illness. It is used in reference to anyobjective evidence of disease that is perceptible to the examiningphysician and/or other healthcare provider.

As used herein, the terms “gastrointestinal” and “GI” refers to thestomach and intestines in the digestive tract of humans and otheranimals. However, as also used in context herein, the term“gastrointestinal tract” (“GI tract”) refers to the entire alimentarycanal, from the oral cavity to the rectum. The term encompasses the tubethat extends from the mouth to the anus, in which the movement ofmuscles and release of hormones and enzymes digest food. Thegastrointestinal tract starts with the mouth and proceeds to theesophagus, stomach, small intestine, large intestine, rectum and,finally, the anus.

As used herein, the term “digestive tract” refers to all of the organsand structures involved in the digestion of nutritional substances(e.g., food and drink), including the oral cavity, esophagus, stomach,small intestine, large intestine, and accessory organs (e.g., the liver,pancreas, etc.).

As used herein, the term “gastrointestinal flora” refers to themicroorganisms that inhabit the gastrointestinal system of humans andother animals. In some particularly preferred embodiments, the term isused in reference to bacterial organisms, but it is not intended thatthe term be so limited.

As used herein, the term “bowel” refers to the intestines.

As used herein, the term “post-prandial” refers to the time after ameal. As used herein, the term “post-prandial symptoms” refers tosymptoms that occur after a subject has ingested a meal.

As used herein, the term “functional bowel disorder(s)” (“FBD”) refersto disorders comprising a variable combination of chronic or recurrentgastrointestinal symptoms that are not explained by structural orbiochemical abnormalities present in the digestive tract of humansand/or other animals.

As used herein, the term “irritable bowel syndrome” (“IBS”) encompassesthe Rome II diagnostic criteria, as known in the art. In particular, theterm is used in reference to at least 12 weeks (which need not beconsecutive), in the preceding 12 months, of abdominal discomfort orpain that has at least two of the following three features: relief withdefecation; and/or onset associated with a change in frequency of stool;and/or onset associated with a change in form (e.g., appearance) ofstool. Symptoms that also support a diagnosis of IBS include: abnormalstool frequency (e.g., greater than 3 bowel movements per day and/orless than 3 bowel movements per week); abnormal stool form (e.g.,lumpy/hard or loose/watery); abnormal stool passage (e.g., straining,urgency, and/or the feeling of incomplete evacuation); passage of mucus;and/or bloating and/or the feeling of abdominal distension.

As used herein, the term “functional abdominal bloating” encompasses theRome II diagnostic criteria, as known in the art. In particular, theterm is used in reference to at least 12 weeks (which need not beconsecutive), in the preceding 12 months, of the feeling of abdominalfullness, bloating and/or visible distension; and insufficient criteriafor a diagnosis of functional dyspepsia, irritable bowel syndrome, oranother functional disorder.

As used herein, the term “functional constipation” encompasses the RomeII diagnostic criteria, as known in the art. In particular, the term isused in reference to at least 12 weeks (which need not be consecutive),in the preceding 12 months, with at least two of the following symptoms:straining during more than ¼ of defecations; lumpy or hard stools formore than ¼ of defecations; the sensation of incomplete evacuation formore than ¼ of defecations; the sensation of anorectalobstruction/blockage during more than ¼ of defecations; requiring manualmaneuvers to facilitate more than ¼ of defecations (e.g., digitalevacuation, support of the pelvic floor); and/or less than 3 defecationsper week.

As used herein, the term “functional diarrhea” encompasses the Rome IIdiagnostic criteria, as known in the art. In particular, the term isused in reference to at least 12 weeks (which need not be consecutive),in the preceding 12 months, during which the subject experiences loose(e.g., mushy) or watery stools that are present more than ¾ of the time,but with no abdominal pain.

As used herein, the term “unspecified functional bowel disorder”encompasses the Rome II diagnostic criteria, as known in the art. Inparticular, it refers to bowel symptoms in the absence of organicdisease that do not fit into the defined categories of functional boweldisorders.

As used herein, the term “relief from gastrointestinal symptoms” refersto reduced or termination of gastrointestinal symptoms, including butnot limited to bloating, abdominal pain, diarrhea, etc.

As used herein, the term “morbidity” refers to illness/disease.

As used herein, the term “mortality” refers to death.

As used herein, the term “incidence” refers to the rate at which acertain event occurs, as in the number of new cases of a specificdisease that occur during a certain period of time.

As used herein, the term “prevalence” refers to the total number ofcases of a specific disease and/or condition in existence in a givenpopulation at a certain time.

As used herein, the term “sequelae” refers to illness/disease andsymptoms/signs that occur as a consequence of a condition and/or diseaseevent. In some embodiments, sequelae occur long after the initialdisease/illness has resolved.

As used herein, the term “sub-clinical infection” refers to infectionthat does not result in the production/observation of signs or symptomsof disease. Often, the patient is infected with a disease-causingorganism, but is unaware of the infection.

As used herein, the term “infection” refers to the invasion andmultiplication of pathogenic microorganisms in the body.

As used herein, the term “culture” refers to any sample or item thatcontains one or more microorganisms. “Pure cultures” are cultures inwhich the organisms present are only of one strain of a particular genusand species. This is in contrast to “mixed cultures,” which are culturesin which more than one genus and/or species of microorganism arepresent. In some embodiments of the present invention, pure culturesfind use. For example, in some particularly preferred embodiments, purecultures of Lactobacillus (e.g., L. acidophilus) find use. However, inalternative embodiments, mixed cultures find use. For example, in someparticularly preferred embodiments, cultures comprised of L. acidophilusand Bifidobacterium find use.

As used herein, the term “Lactobacillus” refers to members of the genusLactobacillus, in the family Lactobacillaceae. These bacteria areGram-positive facultatively anaerobic bacteria that represent a majorpart of the bacterial group often referred to as “lactic acid bacteria.”Various species of Lactobacillus have been identified, including but notlimited to L. acidophilus, L. bulgaricus, L. casei, L. delbrueckii, L.fermentum, L. plantarum, L. reuteri, etc. While it is not intended thatthe present invention be limited to any particular species ofLactobacillus, in some particularly preferred embodiments, L.acidophilus NFCM finds use in the present invention. It is intended thatthe genus include species that have been reclassified (e.g., due tochanges in the speciation of organisms as the result of genetic andother investigations) or renamed for marketing and/or other purposes.

As used herein, the term “Bifidobacterium” refers to members of thegenus Bifidobacterium. These bacteria are Gram-positive anaerobicbacteria that are one of the major strains of bacteria present in thegastrointestinal flora. While it is not intended that the presentinvention be limited to any particular species of Bifidobacterium, insome particularly preferred embodiments, B. lactis Bi-07 finds use inthe present invention. It is intended that the genus include speciesthat have been reclassified (e.g., due to changes in the speciation oforganisms as the result of genetic and other investigations) or renamedfor marketing and/or other purposes.

As used herein, the term “antimicrobial” refers to any compound whichinhibits the growth or kills microorganisms. It is intended that theterm be used in its broadest sense and includes, but is not limited tocompounds such as antibiotics produced naturally or synthetically. It isalso intended that the term encompass compounds and elements that areuseful for inhibiting the growth of or killing microorganisms.

As used herein, the terms “microbiological media,” “culture media,” and“media” refer to any suitable substrate for the growth and reproductionof microorganisms. The term encompasses solid plated media, as well assemi-solid and liquid microbial growth systems.

EXPERIMENTAL

The following example is provided in order to demonstrate and furtherillustrate certain preferred embodiments and aspects of the presentinvention and are not to be construed as limiting the scope thereof.

In the experimental disclosure which follows, the followingabbreviations apply: ° C. (degrees Centigrade); H₂O (water); gm (grams);μg and ug (micrograms); mg (milligrams); ng (nanograms); μl and ul(microliters); ml (milliliters); mm (millimeters); nm (nanometers); μmand um (micrometer); M (molar); mM (millimolar); μM and uM (micromolar);U (units); sec (seconds); min(s) (minute/minutes); hr(s) (hour/hours);sd and SD (standard deviation); PBS (phosphate buffered saline [150 mMNaCl, 10 mM sodium phosphate buffer, pH 7.2]); w/v (weight to volume);v/v (volume to volume); CFU (colony forming units); y/o (years old;years of age); Becton Dickinson (Becton Dickinson Diagnostic Systems,Sparks, Md.); (Difco Laboratories, Detroit, Mich.); GIBCO BRL or GibcoBRL (Life Technologies, Inc., Gaithersburg, Md.); UNC (University ofNorth Carolina); and ATCC (American Type Culture Collection, Manassas,Va.).

The organisms used in the development of the present invention were L.acidophilus NCFM® (PTA-4797) and B. animalis subsp. lactis Bi-07(PTA-4802). These strains were grown at 37° C., in MRS medium (e.g.,Difco, Becton Dickinson) containing 0.05% cysteine. Cultures wereincubated for 48-72 hours in BBL GASPAK™ anaerobic jars containingH₂/CO₂ atmosphere (Becton-Dickinson). For large-scale production, thestrains were grown in fermentation media, harvested by centrifugationand cryostabilized using methods known in the art. The cryostabilizedsolution was lyophilized.

The present invention was assessed using a prospective, randomized,double-blind, placebo controlled study to evaluate the efficacy ofprobiotic (Lactobacillus and Bifidobacterium) bacteria in patients withFBD. The total study duration was 14 weeks. There was a 2 week screeningperiod to determine eligibility, predominant symptoms, and severity atbaseline. This was followed by an 8-week treatment phase, in whichpatients were randomized into 2 treatment arms (i.e., active treatmentwith the probiotic supplement or placebo). The randomization was keptblinded to the patient and the investigators. There was also a 4-weekfollow-up phase after discontinuation of the treatment phase.

Subjects were recruited from the outpatient GI clinic at the Universityof North Carolina Hospitals, from local clinic facilities, and byadvertisement. Power calculations for this study were based on reportedeffects of probiotics on abdominal pain (See e.g., Niedzielin et al.,Eur. J. Gatroenterol. Hepatol., 13:1143-1147 [2001]) since it is afundamental symptom of IBS. Power calculations indicated that the studywas adequately powered.

Subjects meeting all of the following inclusion criteria were determinedto be eligible for this study: signed informed consent was received;18-65 years of age; ambulatory outpatient; presence of IBS, orfunctional diarrhea, or functional bloating according to the Rome IIcriteria for functional GI disorders and the subject has had the abovesymptoms for at least two weeks, despite current therapy (for patientswith diarrhea, the definition of a mean of ≧2 bowel movements per day,or a mean score of ≧4 on the Bristol Stool Form Scale per week wasused); the subject's symptoms were mild to moderate symptoms in severity(symptoms severity was assessed at baseline and at the end of the2-weeks screening period to determine eligibility prior randomization);the subject has had a normal flexible sigmoidoscopy or colonoscopywithin the last 5 years (subjects must have had a colonoscopy if age >50y/o).

Potential subjects were excluded if any of the following criteriaapplied: the subject had inflammation or structural abnormality of thedigestive tract (e.g., inflammatory bowel disease (IBD), duodenal ulcer(DU) or gastric ulcer (GU), obstruction, or symptomatic cholelithiasis);the subject had severe FBD related symptoms at baseline; the subject hasa serious, unstable medical condition; the subject has insulin-dependentdiabetes mellitus; the subject had a major psychiatric diagnosis or asuicide attempt within the last two years; the subject had a history ofalcohol or substance abuse within two years; the subject has abnormallaboratory results (including ALT or AST >than 2.5 times normal, serumcreatinine >2.0 mg/dl, untreated abnormal TSH value); the subject hadbeen treated for a malignancy within the last 5 years (except BCC or SCCskin cancer); the subject had been diagnosed with lactase deficiency andthis could have explained their symptoms (i.e., symptoms resolved orreduced significantly with lactose-free diet); the subject participatedin a drug study within the last 21 days; the subject received antibiotictreatment during the last 8 weeks (for subjects on antibiotic treatment,a washout period of 8 weeks was required); the subject had previoussignificant gastric or intestinal surgery; or the subject was pregnantor lactating, or unwilling to maintain effective contraception duringcourse of the study.

Fifty-seven (57) randomized subjects (females and males) who fulfilledthe Rome II criteria for functional bowel disorders (FBD), IBS, orfunctional diarrhea, and who had at least 6 months of recurrent symptomswere included in this study. Of these, 30 were randomized to the activetreatment group and 27 were assigned to the placebo group. Of theseindividuals, 33 were IBS patients (17 in the active group and 16 in theplacebo group); 21 were FBD patients (non-IBS FBD) (12 in the activegroup and 9 in the placebo group); 3 were functional diarrhea patients(1 in the active group and 2 in the placebo group). The following tableprovides demographic information for each of the individuals in theactive and placebo groups.

TABLE 1 Demographics of Active and Placebo Groups Active Group PlaceboGroup (n = 30) (n = 27) Age (mean; S.D.) 36.0; 10.9 37.0; 14.7 Sex (%female) 72.4 71.4 Education Level (% in each category): Some High School3.3 0.0 High School Graduate 30.0 7.4 Some College or Technical School23.3 40.7 Completed 4 Years of College 20.0 25.9 Some Graduate School23.3 11.1 Completed Graduate Degree 3.3 14.8 Race/Ethnicity: Asian 0.07.4 African-American 16.7 3.7 White 83.3 85.2 Other 0.0 3.7 RelationshipStatus: Single/Never Married 46.7 53.6 Married/Cohabiting 46.7 39.3Separated/Divorced 6.7 7.1

Subjects were seen in the clinic 4 times during this study: (1) at thebeginning of the screening period (week −2); (2) at the beginning of thetreatment phase (week 0); (3) midway through the treatment phase (week4); and (4) at the end of the treatment phase (week 8). Subjects werecontacted by phone two weeks (week 10) and four weeks (week 12) aftercompletion of the treatment phase. In each of the study visits, subjectswere evaluated for the primary and secondary outcome measures using thefollowing tools: GSA (“gastrointestinal symptom-specific anxiety”) DemMed (“major medical descriptors”) (visit 1 only); onset of symptomsquestionnaire (visit 1 only); antibiotic/probiotic treatmentquestionnaire (visit 1 only); satisfaction scale; BDI-II (“BeckDepression Inventory-II); Quality Of Life (IBS-QOL); eating associatedsymptoms questionnaire; IBS severity index; concomitant medications; andreview of adverse events.

During (week 12) and at the end (i.e., week 14) of the follow-up phasesubjects were contacted by phone and asked to mail back their dailydiary cards. Subjects were asked to record their symptoms on diary cardsstarting at the beginning of the screening period and ending 4 weeksafter treatment was discontinued. Subjects recorded the number of bowelmovements, stool consistency for each bowel movement, abdominal pain,bloating, and post-prandial symptoms. In addition, subjects recorded thenumber of pills taken each day. During the phone calls, subjects wereasked about the global relief of their functional GI symptoms, overallwell being, concomitant medications, and about the status of any adverseevents from the study (if there were any). The subjects' symptoms werethen rated on a daily basis.

All of the subjects underwent various laboratory tests, including abreath test to assess for bacterial overgrowth and lactose intoleranceduring the screening period (week −2 to 0). Those with negative resultsobtained during the last 24 months were acceptable; those patients whowere found positive were excluded. Blood tests were conducted toevaluate for treatment safety, at the end of screening and prior tostarting treatment (week 0) (baseline), midway through treatment (week4) and at the end of treatment (8 weeks). This included ESR (erythrocytesedimentation rate) as a marker of inflammation, complete blood countwith platelets and differential (hematocrit, hemoglobin, red blood cellcount, white blood cell count, and differential to include: neutrophils,lymphocytes, monocytes, eosinophils, basophils, bands), sodium,chloride, potassium, calcium, phosphorus, creatinine, BUN, totalprotein, LFTs, albumin, and blood pregnancy test. Laboratory resultsobtained within six months of the screening visit were be allowed to beused as baseline values. The laboratory results were recorded for eachpatient. Although investigation of fecal microflora was an option, thedecision was made not to conduct this aspect of investigation, althoughit is an aspect that could be included in subsequent investigations.

As indicated above, subjects were enrolled into one of two treatmentarms, namely active treatment vs. placebo. The active treatment groupreceived supplements containing L. acidophilus NCFM® and Bifidobacteriumlactis Bi-07. Patients in the active treatment group received oral dosesof equivalent amounts each organism at 1×10¹¹ total CFU (5×10¹⁰ CFU perstrain). The placebo and probiotic products were administered in twiceper day in capsule form, for a total daily dose of 2×10¹¹ CFU.

Throughout the study, the subjects were allowed to maintain theircurrent, stable (≧1 month) medications (these included antispasmodics,fiber supplements, antidiarrheal and/or laxative medications, andantidepressants). However, those patients currently on “anti-gas”medications for their bowel symptoms, were asked to discontinue thesemedications at least 7 days prior to screening visit (i.e., subjectswere not allowed to take these medications within the last week priorentering the screening phase of the study). For those taking antibioticsfor any reason, a washout period of at least 8 weeks was required priorto enrolment in the study. Subjects were also required to remain offthese medications during the remainder of the study. Participants wereasked to keep the same medications to avoid any change in their stablemedications and to avoid taking any medication on an as-needed (i.e.,PRN) basis during study participation. Change in medications during thestudy period was reviewed in each study visit and recorded. In addition,subjects were allowed to consume unintentional probiotics (i.e.,yoghurts). However, they were asked to avoid bacteria-enriched yoghurtsand fermented milks during the study. In addition, probiotic supplementswere not allowed during the study and subjects were asked to discontinuethese supplements at least 8 weeks prior to enrollment in the study.

Data analysis was performed to evaluate significant change in alloutcome measures scores and laboratory tests between pretreatment (i.e.,the end of the screening phase [week 0]), completion of the treatmentphase (week 8), and completion of follow-up period (week 12).Assessments were made for the responsiveness of each of the specificendpoint measures in order to determine the overall responsiveness andthe most specific/positive symptom response. Measure of responsivenessincluded global relief of functional GI symptoms, all specificfunctional GI related symptoms, IBS severity index, and health relatedquality of life. Analysis was conducted for the whole FBD group and, inan exploratory manner, separately for each subgroup (i.e., IBS,functional diarrhea, and non-IBS functional bowel disorder). Thetolerability and safety of the treatment was analyzed with respect tothe incidence of adverse events and changes in laboratory tests. Anupdated log of reported adverse events and laboratory abnormalities foreach patient and for the whole study population was kept.

The results indicated that bloating and distention improvedsignificantly in the probiotics treatment group, as compared to theplacebo group at 4 weeks (4.10±3 vs. 17±3, p=0.009, respectively), andshowed a strong trend of improvement at 8 weeks (4.26±3 vs. 5.84±3,p=0.06, respectively). Secondary analyses using only the IBS subgroup(n=33) showed similar results with significant improvement in bloatingand distention in the probiotics group (n=17), as compared to theplacebo (n=15) group (4.24±3 vs. 6.73±3, p=0.03, respectively). Theseresults indicated that dietary supplements containing L. acidophilusNCFM® and Bifidobacterium lactis Bi-07 (2×10″ CFU total probioticbacteria/day) significantly improved symptoms of bloating and distentionin patients with FBD. Thus, it is contemplated that these probioticswill find use in the management of patients with these disorders.Importantly, it is noted that these probiotics were provided to thesubjects without the addition of other bioactives.

All patents and publications mentioned in the specification areindicative of the levels of those skilled in the art to which theinvention pertains. All patents and publications are herein incorporatedby reference to the same extent as if each individual publication wasspecifically and individually indicated to be incorporated by reference.

Having described the preferred embodiments of the present invention, itwill appear to those ordinarily skilled in the art that variousmodifications may be made to the disclosed embodiments, and that suchmodifications are intended to be within the scope of the presentinvention.

Those of skill in the art readily appreciate that the present inventionis well adapted to carry out the objects and obtain the ends andadvantages mentioned, as well as those inherent therein. Thecompositions and methods described herein are representative ofpreferred embodiments, are exemplary, and are not intended aslimitations on the scope of the invention. It is readily apparent to oneskilled in the art that varying substitutions and modifications may bemade to the invention disclosed herein without departing from the scopeand spirit of the invention.

The invention illustratively described herein suitably may be practicedin the absence of any element or elements, limitation or limitationswhich is not specifically disclosed herein. The terms and expressionswhich have been employed are used as terms of description and not oflimitation, and there is no intention that in the use of such terms andexpressions of excluding any equivalents of the features shown anddescribed or portions thereof, but it is recognized that variousmodifications are possible within the scope of the invention claimed.Thus, it should be understood that although the present invention hasbeen specifically disclosed by preferred embodiments and optionalfeatures, modification and variation of the concepts herein disclosedmay be resorted to by those skilled in the art, and that suchmodifications and variations are considered to be within the scope ofthis invention as defined by the appended claims.

The invention has been described broadly and generically herein. Each ofthe narrower species and subgeneric groupings falling within the genericdisclosure also form part of the invention. This includes the genericdescription of the invention with a proviso or negative limitationremoving any subject matter from the genus, regardless of whether or notthe excised material is specifically recited herein.

What is claimed is:
 1. A method for improving the symptoms of functionalbowel disorder, comprising: i) providing an individual having functionalbowel disorder; ii) providing a composition comprising a probiotic,wherein said probiotic consists of L. acidophilus and Bifidobacteriumlactis; and iii) administering only said composition to said individualunder conditions such that said symptoms of functional bowel disorderare improved, wherein said symptoms of functional bowel disordercomprise bloating and distention.
 2. The method of claim 1, wherein saidcomposition is a dietary supplement.
 3. The method of claim 1, whereinsaid L. acidophilus and Bifidobacterium lactis are administered to saidindividual at a dosage of from about 1×10⁹ to about 2×10¹¹ CFU totalprobiotic bacteria/day.
 4. The method of claim 1, wherein said L.acidophilus and Bifidobacterium lactis are administered to saidindividual at a dosage of about 2×10¹¹ CFU total probiotic bacteria/day.5. The method according to claim 1, wherein the Lactobacillusacidophilus is L. acidophilus NCFM (PTA-4797).
 6. The method accordingto claim 1, wherein the Bifidobacterium lactis is B. animalis subsp.Lactis Bi-07 (PTA-4802).
 7. A method for improving the symptoms offunctional bowel disorder, comprising: i) providing an individual havingfunctional bowel disorder; ii) providing a composition comprising aprobiotic, wherein said probiotic comprises L. acidophilus NCFM(PTA-4797) and B. animalis subsp. lactis Bi-07 (PTA-4802); and iii)administering said composition to said individual under conditions suchthat said symptoms of functional bowel disorder are improved, whereinsaid symptoms of functional bowel disorder comprise bloating anddistention.
 8. A method for improving the symptoms of functional boweldisorder, comprising: i) providing an individual having functional boweldisorder; ii) providing a composition comprising L. acidophilus andBifidobacterium lactis in the absence of additional bioactive compounds;and iii) administering said composition to said individual underconditions such that said symptoms of functional bowel disorder areimproved, wherein said symptoms of functional bowel disorder comprisebloating and distention.